There are currently intensive global research efforts aimed at increasing and modifying the accumulation of lipids, alcohols, hydrocarbons, polysaccharides, and other energy storage compounds in photosynthetic organisms, yeast, and bacteria through genetic engineering. Many improvements have been realized, including increased lipid and carbohydrate production, improved H2 yields, and the diversion of central metabolic intermediates into fungible biofuels. Photosynthetic microorganisms are attracting considerable interest within these efforts due to their relatively high photosynthetic conversion efficiencies, diverse metabolic capabilities, superior growth rates, and ability to store or secrete energy-rich hydrocarbons. Relative to cyanobacteria, eukaryotic microalgae possess several unique metabolic attributes of relevance to biofuel production, including the accumulation of significant quantities of triacylglycerol; the synthesis of storage starch (amylopectin and amylose), which is similar to that found in higher plants; and the ability to efficiently couple photosynthetic electron transport to H2 production. Although the application of genetic engineering to improve energy production phenotypes in eukaryotic microalgae is in its infancy...

Plastid genetic engineering has come of age, becoming today an attractive alternative approach for the expression of foreign genes, as it offers several advantages over nuclear transformants. Significant progress has been made in plastid genetic engineering in tobacco and other Solanaceae plants, through the use of improved regeneration procedures and transformation vectors with efficient promoters and untranslated regions. Many genes encoding for industrially important proteins and vaccines, as well as genes conferring important agronomic traits, have been stably integrated and expressed in the plastid genome. Despite these advances, it remains a challenge to achieve marked levels of plastid transgene expression in non-green tissues. In this review, we summarize the basic requirements of plastid genetic engineering and discuss the current status, limitations, and the potential of plastid transformation for expanding future studies relating to Solanaceae plants.

Phytoremediation to clean up arsenic-contaminated environments has been widely hailed as environmentally friendly and cost effective, and genetic engineering is believed to improve the efficiency and versatility of phytoremediation. Successful genetic engineering requires the thorough understanding of the mechanisms involved in arsenic tolerance and accumulation by natural plant species. Key mechanisms include arsenate reduction, arsenic sequestration in vacuoles of root or shoot, arsenic loading to the xylem, and volatilization through the leaves. Key advances include the identification of arsenic (As) translocation from root to shoot in the As hyperaccumulator, Pteris vittata, and the characterization of related key genes from hyperaccumulator and nonaccumulators. In this paper we have proposed three pathways for genetic engineering: arsenic sequestration in the root, hyperaccumulation of arsenic in aboveground tissues, and phytovolatilization.

In this thesis, I present an engineering discipline for obtaining complex, predictable, and reliable cell behaviors by embedding biochemical logic circuits and programmed intercellular communications into cells. To accomplish this goal, I provide a well-characterized component library, a biocircuit design methodology, and software design tools. I have built and characterized an initial cellular gate library with biochemical gates that implement the NOT, IMPLIES, and AND logic functions in E. coli cells. The logic gates perform computation using DNA-binding proteins, small molecules that interact with these proteins, and segments of DNA that regulate the expression of the proteins. I introduce genetic process engineering, a methodology for modifying the DNA encoding of existing genetic elements to achieve the desired input/output behavior for constructing reliable circuits of significant complexity. I demonstrate the feasibility of digital computation in cells by building several operational in-vivo digital logic circuits, each composed of three gates that have been optimized by genetic process engineering.; (cont.) I also demonstrate engineered intercellular communications with programmed enzymatic activity and chemical diffusions to carry messages...

Explorations and optimizations through the genomic space are a daunting undertaking given the complexity and size of the possible search space. To approach this problem, systematic and combinatorial approaches were employed for the engineering of cellular phenotype in Escherichia coli. Initially, a computational method based on global cellular stoichiometry was employed to identify single and multiple gene knockout targets for lycopene production in E. coli. These targets led to substantial increases in lycopene production, but were limited in scope due to the nature of these models. Therefore, these approaches and targets were complemented with combinatorial searches to identify unknown and regulatory targets. When combined, these searches led to further increases of lycopene production and allowed for the visualization of the resulting metabolic landscape. A more exhaustive search was conducted in the background of eight genotypes which resulted in the formulation of the gene knockout search network. This network enables the investigation into how phenotype optimization is biased by search strategy.; (cont.) Collectively, these results demonstrated that despite the complexity and nonlinearity of genotype-phenotype spaces, most of the significant phenotypes were controlled and regulated by a small subset of key "gateway" nodes. Often...

The successful application of genetic engineering in wheat is dependent on the availability of suitable tissue culture and transformation methods. The primary object of this project was the development of these technologies using elite Australian wheat varieties.; Thesis (Ph.D.)--University of Adelaide, Dept. of Plant Science, Waite Campus, 2001?; Bibliography: leaves 127-151.; xiii, 151, [61] leaves, [19] leaves of plates : ill. (some col.) ; 30 cm.; Title page, contents and abstract only. The complete thesis in print form is available from the University Library.

This review describes the manipulation of the sheep genotype by genetic engineering, which is defined as the introduction or elimination of specific genes in the existing cells of an organism through modern biological techniques for the purpose of changing one or more of its phenotypic characteristics. This can be achieved by direct injection of a DNA sequence for a gene or gene-related function or by transfer of an intact nucleus from a somatic cell containing a set of desirable genes into an enucleated egg cell. The potential of transgenesis over conventional breeding is that single gene changes can be made for a particular phenotype without accompanying multiple gene effects that can be disadvantageous. Several examples of engineered sheep are described. The potential for making other transgenic changes are discussed but transgenic sheep research has virtually ceased because of high cost and the perceived low public acceptance of transgenic animals at the present time.; G. E. Rogers, C. S. Bawden

This paper reports the quantitative and qualitative answers of two groups of public agricultural professionals (a general sample and a targeted sample with some knowledge of organic farming) to issues relating to organic agriculture, genetic engineering, sustainability and associated research issues in Australia. It also analyses what influences these professionals’ views on the sustainability of conventional agriculture in Australia and other agricultural research issues. Professional views towards organic farming and genetic engineering are explored and analysed for their realism. The advent of genetic engineering has been accompanied by growing concern among many of these professionals about safety, public and private research issues, including intellectual property rights, patenting and private funding of public research.; Sarah Ann Wheeler

The aminocoumarin antibiotics novobiocin and clorobiocin are potent inhibitors of bacterial DNA gyrase produced by different Streptomyces strains. Since they are closely related in structure, and their biosynthetic gene clusters have been cloned and sequenced, they represent interesting starting compounds for the development of new antibacterial agents by genetic engineering. However, the functional analysis of biosynthetic genes and the availability of genetic tools for manipulation of producer strains are pre-requisites for such approaches.
Clorobiocin is more potent than novobiocin. It differs from the latter, clinically approved drug in the substitution pattern at C-8´ of the aminocoumarin moiety, carrying a chlorine atom instead of a methyl group, and in the presence of a 5-methylpyrrole-2-carboxyl moiety instead of a carbamoyl group at 3´´-OH of the deoxysugar unit. One aim of this thesis was the production of hybrid antibiotics, combining structural features of these two compounds. By gene inactivation, clo-hal was identified as the gene of the halogenase responsible for the introduction of the chlorine atom of clorobiocin. Expression of the methyltransferase gene novO in the clo-hal- mutant of the clorobiocin producer S. roseochromogenes led to the very efficient formation of a clorobiocin analog with -CH3 instead of -Cl at C-8´ (= novclobiocin 102). However...

Escherichia coli has been widely used to produce high-value recombinant proteins for years. Although high recombinant protein productivity can be attained, perhaps the most important goal for such processes, that of achieving both high gene expression and high cell density simultaneously, is still challenging to both biochemists and biochemical engineers. A series of approaches to overcome this problem are evaluated in this study.
First, a novel pH-inducible gene expression system, in which the expression of foreign gene products is directed by a pH down-shift, was chosen and characterized. This system was shown to have many attractive features, including high-level expression (40% of total cellular protein), fast response, and easy manipulation. It thus can serve as a proper model system for studying the fundamental mechanism of high-level gene expression in E. coli. Second, several factors limiting the culture performance were identified by systematically optimizing the culture conditions. Among those, acetate overproduction was shown to be critically involved. Various approaches on the basis of biochemical and genetic engineering techniques were successfully exploited to bypass such a cultivation bottleneck. Finally, the effects of various genetic elements...

Filamentous bacteriophage (M13) are excellent biological build block due to their multiple peptide display system including type 8 (complete peptide display at pVIII) and type 83 (complete peptide display at both pVIII and pIII) display systems. Unlike the phagemid systems, the advantage of these systems is that we can get homogenous peptide display on pVIII resulting in uniform placement of selected molecules as well as defined length and width. In this thesis, type 8 and type 83 phage were constructed and used as biological scaffolds to meet the following four specific aims. First, the self-assembly of engineered M13 bacteriophage as a template for Co-Pt crystals was demonstrated. An phage library with an octapeptide library on the major coat protein (pVIII) was used for selection of binders to cobalt ions. Fibrous structures with directionally ordered phage were obtained by interaction with cobalt ions. Co-Pt alloys were synthesized on the fibrous scaffold, and their magnetic properties were characterized. The mineralization showed organized nanoparticles on fibrous bundles with superparamagnetic properties. Second, an in vitro molecular selection method in non-biological conditions for inorganic synthesis was introduced.; (cont.) A phage display peptide library which is resistant to ethanol was constructed and used for selection against titania in 90% ethanol. The selected peptide...

This is the final version. It was first published by NPG at http://www.nature.com/srep/2015/150605/srep11061/full/srep11061.html.; The accelerated discovery of disease-related genes emerging from genomic studies has strained the capacity of traditional genetically engineered mouse models (GEMMs) to provide in-vivo validation. Direct, somatic, genetic engineering approaches allow for accelerated and flexible genetic manipulation and represent an attractive alternative to GEMMs. In this study we investigated the feasibility, safety and efficiency of a minimally invasive, lentiviral based approach for the sustained in-vivo modification of renal tubular epithelial cells. Using ultrasound guidance, reporter vectors were directly injected into the mouse renal parenchyma. We observed transgene expression confined to the renal cortex (specifically proximal and distal tubules) and sustained beyond 2 months post injection. Furthermore, we demonstrate the ability of this methodology to induce long-term, in-vivo knockdown of candidate genes either through somatic recombination of floxed alleles or by direct delivery of specific shRNA sequences. This study demonstrates that ultrasound-guided injection of lentiviral vectors provides a safe and efficient method for the genetic manipulation of renal tubules...

This dissertation explores philosophical problems in biology, particularly those relating to macroevolutionary theory. It is comprised of a series of three papers drawn from work that is currently at the publication, re-submission, and review stage of the journal refereeing process, respectively. The first two chapters concern the overarching contours of complex life, while the third zeroes in on the short and long-term prospects of human evolution.

The rhetorical journey begins with a thought experiment proposed by the late paleontologist Stephen Jay Gould. Gould hypothesized that replaying the "tape of life" would result in radically different evolutionary outcomes, both with respect to animal life in general and the human species in particular. Increasingly, however, biologists and philosophers are pointing to convergent evolution as evidence for replicability and predictability in macroevolution. Chapters 1 and 2 are dedicated to fleshing out the Gouldian view of life and its antithesis, clarifying core concepts of the debate (including contingency, convergence, constraint and causation), and interpreting the empirical data in light of these conceptual clarifications. Chapter 3 examines the evolutionary biological future of the human species...

Disruption of coordinated impulse propagation in the heart as a result of fibrosis or myocardial infarction can create an asynchronous substrate with poor conduction and impaired contractility. This can ultimately lead to cardiac failure and make the heart more vulnerable to life-threatening arrhythmias and sudden cardiac death. The transplantation of exogenous cells into the diseased myocardium, "cardiac cell therapy," has been proposed as a treatment option to improve compromised cardiac function. Clinical trials of stem cell-based cardiac therapy have shown promising results, but also raised concerns about our inability to predict or control the fate of implanted cells and the electrical consequences of their interactions with host cardiomyocytes. Alternatively, genetically engineered somatic cells could be implanted to selectively and safely modify the cardiac electrical substrate, but their unexcitable nature makes them incapable of electrically repairing large conduction defects. The objective of this thesis was thus to develop a methodology to generate actively conducting excitable cells from an unexcitable somatic cell source and to demonstrate their utility for studies of basic electrophysiology and cardiac cell therapy.

This paper deals with the progress made in genetic engineering techniques, capable of altering the genetic potential of an organism, either by the introduction or the suppression of new structural genes. Some of the general applications are described as are also, more particularly, their uses in the field of medicine. A critical analysis of the benefits and risks involved is also undertaken.; São abordados os progressos havidos com as técnicas de engenharia genética, capazes de alterar o potencial genético de um organismo, quer pela introdução, quer pela supressão de novos genes estruturais. São mencionadas algumas das aplicações em geral e, em particular, possibilidades de uso no campo da medicina. É feita uma análise crítica dos benefícios e riscos envolvidos.

In this article, the concept of"liberal democracy" and its implications for biotechnology such as human genetic engineering will be examined. Liberal democracy is characterised by a number of features, some of which has extensive repercussions on biotechnology, especially concepts such as the equal protection of human rights, civil rights, civil liberties, political freedom for all people and autonomy and libertarianism. Advocates of human genetic engineering for purposes other than the healing of genetically transmitted diseases often appeal to these features in the quest for its legalisation. I will examine whether the attributes of liberal democracy would indeed justify the use of this type of biotechnology and if yes, what a possible theological response would be, drawing on the political theology of Jürgen Moltmann.